Isolation of Chloroplast Thylakoid Membranes

The thylakoid membrane of the chloroplast is an internal interconnected membrane system in which most of the photosynthesis-related proteins and protein complexes are localized in the thylakoid membrane and thylakoid lumen. The thylakoid membrane has always been a hotspot in photosynthesis research. Lifeasible's expertise in chloroplast engineering has allowed us to isolate a fully functional thylakoid membrane section through advanced techniques and gain a deeper understanding of chloroplast thylakoid function and its metabolic laws. If you have any special requirements about our isolation of chloroplast thylakoid membranes services, please feel free to contact us. We are looking forward to working together with your attractive projects.

Chloroplast Thylakoid Membranes

Plant chloroplast thylakoid is the site of light reaction in photosynthesis, and is the site of light capture, photosynthetic electron transfer, proton translocation and ATP synthesis. Therefore, most of the proteins and protein complexes related to photosynthesis are located in thylakoid Membranes and thylakoid lumen. The thylakoid membrane system mainly includes four highly abundant multi-subunit membrane protein complexes, namely the photosystem I (PSI), PSII, ATP-synthase and cytochrome b/f complexes. These protein complexes are key sites for light energy absorption, transmission and conversion in plants. With the continuous improvement and development of plastid extraction and separation techniques, much understanding of the photosynthesis process has become possible by studying the structure and function of isolated thylakoids, thylakoid membranes and protein complexes. In addition, researchers have successfully used two-dimensional isoelectric focusing/sodium dodecyl sulfate-polyacrylamide gel electrophoresis (2D-IEF/SDS- PAGE) to separate Arabidopsis chloroplast luminal proteins, pea peripheral thylakoid proteins, and chloroplasts protein. The premise of the research on thylakoid protein is to isolate the intact and highly purified chloroplast thylakoid membrane.

Fig. 1. Schematic overview of the light-dependent de-etiolation process and the biogenesis of thylakoid membranes. (Pribil M, et al., 2014)

Services

Lifeasible is committed to developing reliable methods to isolate functional thylakoid membranes. Just as the best intact chloroplasts are isolated by osmotic rupture of protoplasts, we also obtain the best thylakoids from intact chloroplasts. This approach not only achieves that the thylakoid membrane is protected by the chloroplast envelope from degradation by proteases and lipases present in the cell's chloroplast lumen. In addition, non-specific adsorption of non-chloroplast proteins and nucleic acids to the thylakoid membrane surface is avoided. Therefore, we can help you easily prepare intact and highly purified chloroplast thylakoid membranes.

However, isolation of thylakoids from intact chloroplasts is sometimes limited due to small yields. Therefore, our scientists used a more direct sequestration process to obtain chloroplast thylakoid membranes. This method yields pure, structurally intact and functional thylakoid membranes from most species commonly found in the laboratory.

During photosynthesis, electron transport and photophosphorylation occur in thylakoid membranes. Despite the need to understand the organizational relationship between light- and dark-responsive components in the body, research into photosynthesis has focused on studying these components individually. The supramolecular organization of soluble proteins and enzymes around the thylakoid membrane remains to be studied. Therefore, we performed ultracentrifugation after obtaining the thylakoid membrane to further isolate the thylakoid membrane components with photosynthetic functions, including all components with electron transport chain, ATP synthase, phycobiliprotein, ferredoxin-NADP reductase.

Competitive Advantages

• The leaves are cut into small pieces before separation and macerated to make separation faster and more efficient.
• The thylakoid membrane retains its original function.
• The thylakoid membrane structure is intact, maintaining the distinction between stacked and non-stacked regions.
• Choosing reasonable methods to reduce the contamination of substrates and extra-chloroplasts.
• To meet the special purpose of the isolated thylakoid.

References

1. Pribil M, Labs M, Leister D. (2014) Structure and dynamics of thylakoids in land plants[J]. Journal of experimental botany. 65(8): 1955-1972.
2. Andersson B, Anderson J M. (1985) The chloroplast thylakoid membrane—isolation, subfractionation and purification of its supramolecular complexes[M]//Cell Components. Springer, Berlin, Heidelberg. 231-258.